CAJ | 학술논문

目的采用SYBR GreenⅠ荧光定量聚合酶链反应方法,研究颈7神经根切断后靶肌肉内乙酰胆碱受体γ-和ε-亚基的mRNA表达水平的变化,来探索颈7神经根切断后的代偿机制.方法 36只雄性SD大鼠,随机分成6组,每组6只.右侧建成颈7神经根切断模型,左侧为健康对照组.用SYBR GreenⅠ荧光定量RT-PCR方法,检测术后第1、2、4、6、8、12周肱三头肌内乙酰胆碱受体γ-、ε-和α-亚基的mRNA表达水平的变化曲线.收集所有数据后,用SPSS12.0软件进行均值、标准差计算和方差统计学分析.结果 ε-AchR的mRNA在颈7神经根切断术后第1～6周下调表达(P＜0.05),第6周下降到谷底,在第12周基本恢复到健康对照组水平.γ-AchR的mRNA表达在颈7神经根切断术后第1周就开始上升,到第6周达顶峰(P＜0.05),在第12周下降至正常水平.结论运用SYBRGreenⅠ荧光RT-PCR技术,证实在SD大鼠颈7神经根切断后第1～6周内,其靶肌肉内γ-AchR/ε-AchR会出现失神经的转换,但在第12周左右恢复正常表达.
목적 채용SYBR GreenⅠ형광정량취합매련반응방법,연구경7신경근절단후파기육내을선담감수체γ-화ε-아기적mRNA표체수평적변화,래탐색경7신경근절단후적대상궤제.방법 36지웅성SD대서,수궤분성6조,매조6지.우측건성경7신경근절단모형,좌측위건강대조조.용SYBR GreenⅠ형광정량RT-PCR방법,검측술후제1、2、4、6、8、12주굉삼두기내을선담감수체γ-、ε-화α-아기적mRNA표체수평적변화곡선.수집소유수거후,용SPSS12.0연건진행균치、표준차계산화방차통계학분석.결과 ε-AchR적mRNA재경7신경근절단술후제1～6주하조표체(P＜0.05),제6주하강도곡저,재제12주기본회복도건강대조조수평.γ-AchR적mRNA표체재경7신경근절단술후제1주취개시상승,도제6주체정봉(P＜0.05),재제12주하강지정상수평.결론 운용SYBRGreenⅠ형광RT-PCR기술,증실재SD대서경7신경근절단후제1～6주내,기파기육내γ-AchR/ε-AchR회출현실신경적전환,단재제12주좌우회복정상표체.
Objective To explore the compensatory mechanisms after transection of C7 nerve root by quantifying ε-AchR/γ-AchR subunit mRNA expression using RT-PCR with SYBR Green Ⅰ. Methods Thirtysix male SD rats were randomly divided into 6 groups, with 6 each. The right C7 nerve root was transected while the left C7 was left intact to serve as control. mRNA expressions of ε-AchR/γ-AchR subumits in the triceps brachii muscles of experimental sides and normal sides were tested at week 1, 2, 4, 6, 8, and 12 after G7 nerve root transection using real-time fluorescence quantitative revere transcriptase-polymerase chain reaction with SYBR Green Ⅰ. SPSS12.0 software was used to calculate the mean and standard deviation of the values, and statistical analysis of the variance. Results In the 1st, 2nd, 4th, and 6th week, mRNA expression level of ε-AchR subunit of the experimental group was down-reg ulated comparing to that of the control( P ＜ 0.05). The level of ε-AchR subunit mRNA expression was lowest during the 6th week, but then returned to normal level 12 weeks after the transection. γ-AchR subunit mRNA expression increased in the first postoperative week and peaked during the 6th week ( P ＜ 0.05). γ-AchR subunit mRNA expression retuned to normal level 12 weeks after the transection. Conclusion The change of ε-AchR/γ-AchR subunit mRNA expression indicated that C7 nerve root transection caused denervation changes in subtype structures of the target muscle within the first 6 weeks.Compensatory recovery occurred in the 12th week.